Separation of Plant Pigments Using Paper Chromatography

Paper chromatography is a useful technique for separating and identifying pigments and other molecules from cell extracts that contain a complex mixture of molecules. The solvent moves up the paper by capillary action, which occurs as a result of the attraction of solvent molecules to the paper and the attraction of solvent molecules to one another. As the solvent moves up the paper, it carries along any substances dissolved in it. The pigments are carried along at different rates because they are not equally soluble in the solvent and because they are attracted, to different degrees, to the fibers in the paper through the formation of intermolecular bonds, such as hydrogen bonds.

Beta carotene, the most abundant carotene in plants, is carried along near the solvent front because it is very soluble in the solvent being used and because it forms no hydrogen bonds with cellulose. Another pigment, xanthophylls, differs from carotene in that it contains oxygen. Xanthophyll is found further from the solvent front because it is less soluble in the solvent and has been slowed down by hydrogen bonding to the cellulose. Chlorophylls contain oxygen and nitrogen and are bound more tightly to the paper than are the other pigments. Chlorophyll a is the primary photosynthetic pigment in plants. a molecule of chlorophyll a is located at the reaction center of photosystems. Other chlorophyll a molecules, chlorophyll b, and the carotenoids (that is, carotenes and xanthophylls) capture light energy and transfer it to the chlorophyll a at the reaction center. Carotenoids also protect the photosynthetic system from the damaging effects of ultraviolet light.